pAAVLINKv1-pCALM1-Cre-3'-NCKAP5-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-NCKAP5-FLAG                7153 bp DNA     circular SYN 18-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    12..130
                     /label=AAV2 ITR
     promoter        145..265
                     /label=pCALM1
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          1309..1349
                     /label=Splicing Donor
     protein_bind    1356..1389
                     /label=lox2272-JTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          1390..1481
                     /label=Splicing Acceptor
                     /note="Splicing acceptor"
     CDS             1482..3866
                     /codon_start=1
                     /label=NCKAP5
                     /translation="VSSSSSSSSPAKSHNSPHGCQSAHEKGLKTRLPVGLKVLMKSPQL
                     LRKSSTVPGKHEKDSLNEASKSSVAVNKSKPEDSKNPASMEITAGERNVTLPDSQAQGS
                     LADGLPLETALQEPLESSIPGSDGRDGVDNRSMRRSLSSSKPHLKPALGMNGAKARSHS
                     FSTHSGDKPSTPPIEGSGKVRTQIITNTAERGNSLTRQNSSTESSPNKAPSAPMLESLP
                     SVGRPSGHPSSGKGSLGSSGSFSSQHGSPSKLPLRIPPKSEGLLIPPGKEDQQAFTQGE
                     CPSANVAVLGEPGSDRRSCPPTPTDCPEALQSPGRTQHPSTFETSSTSKLETSGRHPDA
                     SATATDAVSSEAPLSPTIEEKVMLCIQENVEKGQVQTKPTSVEAKQKPGPSFASWFGFR
                     KSRLPALSSRKMDISKTKVEKKDAKVLGFGNRQLKSERKKEKKKPELQCETENELIKDT
                     KSADNPDGGLQSKNNRRTPQDIYNQLKIEPRNRHSPVACSTKDTFMTELLNRVDKKAAP
                     QTESGSSNASCRNVLKGSSQGSCLIGSSISTQGNHKKNMKIKADMEVPKDSLVKEANEN
                     LQEDEDDAVADSVFQSHIIESNCQMRTLDSGIGTFPLPDSGNRSTGRYLCQPDSPEDAE
                     PLLPLQSALSAVSSMRAQTLEREVPSSTDGQRPADSAIVHSTSDPIMTARGMRPLQSRL
                     PKPASSGKVSSQKQNEAEPRPQTCSSFGYAEDPMASQPLPDWGSEVAATGTQDKAPRMC
                     TYSASGGSNSDSDLDYGDNGFGAGRGQLVKALKSAAPEIETT"
     misc_feature    2173..2178
                     /label=SDS
     CDS             3873..3938
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3942..4373
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    4086..4091
                     /label=SDS
     repeat_region   4416..4556
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4416..4545
                     /label=AAV2 ITR
     rep_origin      4631..5086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5368..5472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5473..6333
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6504..7092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtcagcagt tcctcatcat
     1501 cctcatcacc cgccaaaagc cataacagcc ctcatggttg tcaaagtgct catgagaaag
     1561 gactgaaaac tcgccttcca gtgggactca aagtgctcat gaagtctccc cagctgctca
     1621 ggaaaagttc caccgtgcca gggaaacatg aaaaagacag tttaaatgaa gcctccaaaa
     1681 gttccgtggc tgtgaacaag tctaagccag aggactccaa gaatccagca agcatggaga
     1741 tcacagcggg tgaaagaaat gtgaccctac cggattcaca agcacagggc agtttagctg
     1801 atgggcttcc cctggaaaca gcactacaag agccattgga aagtagcatc cctgggagtg
     1861 atggaaggga tggggtagat aatagatcca tgagaagatc cctttcctcc agcaaaccac
     1921 acctaaaacc agctctgggt atgaatggcg ccaaagcccg cagccacagc ttcagtacac
     1981 actcaggaga caagccttct acgcccccca tcgaagggtc aggcaaagtc cgcactcaga
     2041 tcattaccaa taccgccgag agaggcaatt ctcttacccg gcagaactct tccacggaaa
     2101 gctctcccaa caaggcccct tctgctccca tgttggagag tctccccagt gttgggaggc
     2161 cctcggggca cccctcctcc gggaagggct ccctggggag ctcaggcagc ttcagcagtc
     2221 agcatgggag cccaagtaag ttgcctttga ggatccctcc aaagtctgag ggactcctca
     2281 tcccacctgg aaaggaagac cagcaggcct tcacccaggg agagtgcccc agtgccaatg
     2341 tggctgtact tggggaacca ggcagtgacc gccgcagttg cccacccacc ccaacagact
     2401 gccctgaagc cctgcagagc ccagggagga ctcagcatcc aagcactttt gaaacaagca
     2461 gtacatccaa gctagaaact tctggaaggc atccagatgc ctctgcaacc gcgactgatg
     2521 ctgtgagttc agaagccccc ctctcaccca caatcgaaga aaaggtcatg ttgtgcattc
     2581 aggaaaatgt ggaaaagggc caagtgcaaa caaagcccac ctctgtggaa gcaaagcaga
     2641 agcctgggcc ttcttttgcc agctggtttg gttttcggaa gagtagactt ccagctctga
     2701 gtagcaggaa aatggacatc tccaaaacca aagtagaaaa gaaagatgca aaagtcttgg
     2761 ggtttggaaa cagacaacta aaatcagaaa gaaaaaaaga gaaaaagaag cctgaactac
     2821 agtgtgagac agaaaatgag cttatcaagg acaccaagtc agcagataat ccagatggcg
     2881 gtttacaaag caaaaataat cggagaacac cacaagacat ttacaaccaa ctgaagattg
     2941 aaccaaggaa tagacacagc cctgttgcat gttcaacgaa agacaccttc atgacggaac
     3001 tcttgaacag agttgataag aaagcagctc cacagacaga aagtggatca agtaatgctt
     3061 cctgcaggaa tgtgttaaag ggcagttctc agggctcctg tctcatcggc agctctatca
     3121 gtactcaagg aaaccacaag aaaaacatga aaatcaaagc cgatatggaa gtaccaaaag
     3181 actccctggt aaaagaggca aatgaaaact tgcaagagga tgaagacgat gcagttgcag
     3241 attctgtatt tcagagccac atcatagaat ccaactgcca gatgagaaca ttggacagtg
     3301 ggatcggaac ctttccactc ccagactcgg gaaatcgctc gacaggacgc tacctatgcc
     3361 agccagactc cccagaggac gctgagcctc tcctgcctct ccagtcagcc ctttctgcag
     3421 tttcttccat gagagcccaa acccttgaac gtgaagtgcc ttcctccaca gacggccagc
     3481 gccctgcaga tagcgccatt gttcattcca catccgaccc catcatgacc gccagaggga
     3541 tgaggcctct tcagagccgc ctccccaaac cagcttcctc aggaaaagtc agttcccaaa
     3601 agcagaatga agcagagcca aggcctcaga catgctcatc attcggatat gctgaagacc
     3661 caatggcaag ccagccgctt ccagactggg ggagtgaagt tgctgccacc gggacccagg
     3721 acaaggcacc cagaatgtgt acgtactctg ccagcggtgg cagtaatagt gacagtgacc
     3781 tggactatgg agataatggt tttggagctg gaaggggaca gttagtgaaa gcactgaaga
     3841 gcgctgcccc agaaattgag acaacttgta cagactacaa agaccatgac ggtgattata
     3901 aagatcatga catcgactac aaggatgacg atgacaagta actcgagata atcaacctct
     3961 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4021 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4081 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     4141 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgtttat
     4201 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt
     4261 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4321 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagatctg
     4381 gagaccggtg tggcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4441 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4501 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4561 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4621 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4681 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     4741 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4801 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4861 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4921 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4981 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     5041 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5101 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5161 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5221 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5281 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5341 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5401 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5461 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5521 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5581 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5641 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5701 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5761 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5821 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5881 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5941 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6001 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6061 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6121 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6181 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6241 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6301 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7141 tttgctcaca tgt
//